The invention herein relates to a solution, which, in addition to the standard examinations of the eye fundus, allows for mapping and/or examination of the photoreceptors in the human eye in order to diagnose and treat diseases at an early stage. Changes of those light-sensitive sensory cells of an eye, called photoreceptors, are frequently the first signs of diseases. Thereby, an early detection drastically increases the chances for a successful treatment.
The human eye exhibits, for example, two types of photoreceptors: Rods and cones. While the light-sensitive rods allow for night vision, the cones allow for daylight vision and color sensitivity (red, blue, green).
The examination and evaluation of pathological changes of the visible part of the eye, particularly the retina and its blood-supplying vessels, are called ophthalmoscopy or funduscopy. Thereby, the vessels, which supply the retina with blood, are clearly distinguishable from the retina, whereby the light-red appearing arteries, which originate in the optic nerve papilla (blind spot), are also distinguishable from the dark-red appearing veins.
In ophthalmoscopy, two different methods are commonly applied. While the so-called direct ophthalmoscopy, whereby the ophthalmoscope is placed at a distance of approximately 10 cm, i.e., directly in front of the patient's eye, and achieves an angle of view of 10 to 15° with a magnification of up to 15 times, is better suited for examining details, such as optic nerve papilla, vessel origins, and the yellow spot (macula lutea), indirect ophthalmoscopy at a distance of approximately 50 cm with a magnification of 2 to 5 times and an angle of view of 25 to 40°, is better suited for the viewing and examination of the eye fundus as a whole. Most eye specialists prefer the indirect ophthalmoscopy because it provides a significantly better overview due to the smaller magnification and, unlike direct ophthalmoscopy, it allows for a stereoscopic (3D) evaluation. Furthermore, indirect ophthalmoscopy can also be performed with the slit lamp, known as a standard device for the examination of the eye by eye specialists. With a slit lamp the retinal image can be magnified and/or evaluated with an even greater 3D effect through projection of a slit of light. However, examination with a slit lamp is not suitable for observing details in the retina due to insufficient resolution of the lens system of a slit lamp.
In addition to ophthalmoscopy, OCT assemblies (Optical Coherence Tomography) are now commonly used for detailed optical examination of the eye fundus. OCT is an examination method, whereby temporally short coherent light is utilized with the help of an interferometer for distance measurement of the reflective materials to be found in the eye.
The basic principle of OCT is based on white light interferometry, whereby the durations of a signal are compared with each other with the help of an interferometer (most commonly, a Michelson interferometer). Thereby, one arm of the interferometer with known optical path length (=reference arm) is used as reference for the measurement arm.
The interference of the signals (optical cross correlation) from both arms results in an interference pattern from which the relative optical path length can be determined within an A-scan (individual depth signal). Subsequently, the beam in the one-dimensional raster scan is led transversally in one or two directions, whereby a two-dimensional B-scan or a three-dimensional tomogram (C-scan) can be obtained.
The preeminent feature of OCT is the decoupling of the transversal from the longitudinal resolution. In conventional light microscopy, the axial resolution (depth) as well as the transversal resolution depends on the focusing of the light beam. The parameter for the focusability is the numerical aperture. In OCT, the resolution is only restricted by the bandwidth of the applied light. Therefore, with great bandwidth (wide spectrum), a high resolution, with which small details can be resolved, is achievable.
The areas of application for OCT are primarily in medicine, particularly, ophthalmology, as well as for early cancer diagnosis and skin examinations. Thereby, reflections are measured at boundaries of materials with varying refractive indices and, subsequently, a three-dimensional image is reconstructed. Such a reconstruction is called tomography.
Currently, the main application is the examination of the eye fundus and/or the posterior eye segment since competing technologies, such as the confocal microscope, can only insufficiently map the fine layer structure of the retina with its thickness of approximately 250-300 μm due to the small size of the pupil and the great distance between cornea and retina. Most of all, the significant advantage of OCT is the contact-free measurement since risks of infection and emotional stress are largely avoided.
A further commonly used method for detailed optical examination of the eye fundus is fundus autofluorescence (FAF). With this method, lipofuscin accumulation can non-invasively be detected in vivo in the lysosomal compartment of the single-layered retinal pigment epithelium (RPE).
In [1], A. Bindewald et al. describe a further developed method for scanning laser ophthalmoscopy.
Thereby, resolutions of up to 5 μm/pixel are achieved with the use of confocal scanning laser ophthalmoscopes (cSLO) on the basis of solid-state lasers for generating excitation laser light (488 nm), causing changes of the topographical FAF intensity distribution to appear in different retinal pathologies, including age-related macular degeneration, macular edema, and genetically determined retinopathies. Internal fixation control, magnification of the focus area, improved lens system, and a new laser source result in advantages for clinical applications. Improved quality of FAF images with the new cSLO is of importance for clinical diagnostics and the precise phenotyping of retina diseases for scientific purposes as well as for future therapy monitoring on RPE cell basis.
With the methodical advancement of confocal scanning laser ophthalmoscopy, FAF images of hitherto unknown quality are produced, which even allows for the differentiation of individual RPE cells in vivo. Aside from reliable findings with prior applications, possibilities for new therapeutic methods also arise.
The disadvantages of both methods lie in the facts that they are not based on standard devices commonly used by eye specialists as well as their extremely high costs.
The slit lamp as well as the fundus camera are considered conventional standard devices for eye specialists. While the slit lamp more commonly applies to the examination of the anterior eye segments, the fundus camera is designated for the examination of the eye fundus.
Thereby, fundus cameras, which are equipped with image registration units for documentation, are commonly used. Pictorial representation of the retina of the human eye is an important aid for diagnoses. However, the technical realization of images of the eye fundus is no trivial matter due to the optical structure of the eye.
Thereby, new methods forgo the use of pupil-dilating measures on the patient and work with infrared illumination. The quality of the findings images essentially depends on the optical positioning of the fundus camera, however, the optical properties of the eye itself, as part of the imaging optical path, limit the achievable results. Advanced fundus cameras are equipped not only with a digital imaging unit but also with image processing and archiving systems. For the examination of the eye fundus with fluorescent solutions, which are added to the patient's blood, applicable excitation and band-elimination filters are present in the optical path, which are swiveled into the optical path, if needed.
Since the viewing angle of classic fundus cameras is approximately 60°, detailed examinations of the photoreceptors in the human eye are not possible because the resolution of their lens systems is insufficient for said purpose.
By means of optical adjustments, the conventional viewing angles of 45 to 60° for fundus imaging could be decreased, thereby allowing for mapping of smaller segments of the human retina with corresponding higher resolution. But the structures of the photoreceptors cannot be made visible even with higher resolution because the depth of field of the fundus camera is too low. In order to map the photoreceptors of the human retina, transverse resolutions of less than 5 μm are required. Furthermore, the additionally mapped out-of-focus layers of the retina during fundus imaging with a fundus camera have further adverse effect on the imaging quality.
In [2] and WO 1996/24082 A1, Gustafsson describes that in a fluorescence microscope the lateral resolution can be magnified by a factor of two by illuminating the sample with a spatially structured light source.
Through illumination with a series of excitation patterns, high-resolution information, which is usually inaccessible, is encoded into the observed image. The stored images are processed linearly in order to extract the new information and produce a reconstruction with double resolution. Unlike confocal microscopy, the entire emission light is used, producing images with greater clarity when compared to confocal microscopy.
According to prior art, solutions are also known, whereby the sample to be examined is illuminated with periodic patterns in order to achieve an increase in resolution.
The solution described in U.S. Pat. No. 5,867,604 A relates to a system for the improvement of the resolution of imaging systems by means of illumination with a periodic pattern. Thereby, particularly the illumination phase is altered in order to extract the amplitude and the phase data from the received scattered light images and produce synthetic three-dimensional images. The periodic illumination patterns can be produced from diffraction gratings as well as by interferometric means. In a particularly advantageous embodiment, the application of the described method in a confocal microscope is described.
WO 1998/045745 A1 also describes a solution which relates to a system for the improvement of the resolution of imaging systems by means of illumination with periodic patterns. Thereby, the illumination pattern is moved continuously or discretely, so that at least three images of the sample can be produced with varying phasing of the illumination pattern. An evaluation unit removes the spatial patterns from the images, resulting in an image of the sample which is optically divided into sections. In a particularly advantageous embodiment, the application of the described method in a conventional microscope is described.
Even though both descriptions contain references regarding the application of the solutions for other optical imaging systems, a specific application, as, for example, in ophthalmology, is not mentioned.
In U.S. Pat. No. 5,116,115 A, a method and an assembly for measuring the topography of the cornea of an eye is described. Thereby, a thin, flexible, reflecting material is placed in such a way on the cornea to be examined that it adjusts exactly to the shape of the cornea. As a result, it is possible to capture the shape of the cornea by means of the projected pattern. For said purpose, structured sinusoidal patterns with varying phasings are projected onto the cornea and mapped by a detector. A computer calculates an elevation map from the digitalized images at various phasings. By means of the calculated elevation map, the topography of the cornea can be viewed. The basic idea of this described solution consists of the determination of the topography of the cornea of an eye. Once again, no references are disclosed or implied with regard to other applications for the eye.
Methods and assemblies for microscopic imaging, whereby objects are illuminated from a light source with periodic patterns, are described in U.S. Pat. No. 6,376,818 B1 and EP 1 412 804 B1. Thereby, the periodic pattern consists of a striped pattern. By means of a microscope, at least three images of the object are mapped at varying spatial phasings and transmitted to an evaluation unit. From the analysis of those three images, a three-dimensional image of the volume structure of the object is derived, which generally only contains in-focus details. With the suggested solution, an assembly and method for producing three-dimensional images of an object to be examined are provided, which, similar to the confocal images, essentially only contain in-focus details.
Even though the description contains a reference regarding, particularly, the biomedical application of the solution, a specific application as, for example, in ophthalmology, is not mentioned.
In [3], Douglas Starkey describes an optical projection system for the detection of glaucoma diseases. In this solution, modifiable patterns, produced by an interferometer, are projected onto the retina of the eye to be examined. The electric potentials, which result from light impinging on the retina, are recorded with the help of electrodes as electroretinogram (ERG). The ERG shows the sum of the responses of the entire retina. The projection of certain patterns onto the retina allows, particularly, for the evaluation of the inner layers of the retina. Thereto, Starkey developed a device with which sinusoidal patterns are produced by means of a laser interferometer and projected onto the retina. The thereby recorded electroretinogram is also called PERG (pattern electroretinogram). While previously applied solutions were depicted by means of projectors or TV screens, the patterns in Starkey's solution are produced by a laser interferometer, including changes with regard to contrast, intensity, and spatial/temporal frequency. This allows for a significant reduction of distortions as well as chromatic deviations.
The modifiable patterns produced by Starkey are used exclusively for producing electroretinograms, particularly, PERG. No references are disclosed or implied with regard to other applications for the eye.